DNA methylation in mouse A-repeats in DNA methyltransferase-knockout ES cells and in normal cells determined by bisulfite genomic sequencing.
Mouse ES cells with a null mutation of the known DNA methyltransferase retain some residual DNA methylation and can methylate foreign sequences de novo. We have used bisulfite genomic sequencing to examine the sequence specificity and distributions of methylation of a hypermethylated CG island sequence, mouse A-repeats. There were 13 CG dinucleotides in the region examined, 12 of which were methylated to variable extents in all DNAs. We found that: (1) there is considerable residual DNA methylation in ES cells lacking the known DNA methyltransferase (29% of normal methylation in the complete knockout ES DNA); (2) this other activity methylates at exactly the same CG sites as the major methyltransferase; and (3) differences in the distribution of methylated sites between A-repeats in these DNAs are consistent with this other activity methylating in a random de novo fashion. Also, the lack of any methylation in non-CG sites argues that, in other studies where non-CG methylation sites have been found by bisulfite sequencing, detection of such sites of non-CG methylation is not an inherent artifact in this methodology.
Woodcock
D M
DM
Peter MacCallum Cancer Institute, Melbourne, Victoria, Australia. woodcock@petermac.unimelb.edu.au
Linsenmeyer
M E
ME
Warren
W D
WD
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D018899:CpG Islands; D004247:DNA / Q000032:analysis; D019175:DNA Methylation; D015254:DNA Modification Methylases / Q000235:genetics* / Q000378:metabolism; D017931:DNA Primers; D018900:Dinucleotide Repeats; D017353:Gene Deletion; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA; D013447:Sulfites
D017931:DNA Primers; D013447:Sulfites; D004247:DNA; D015254:DNA Modification Methylases; C042345:hydrogen sulfite